Consensus protein engineering on the thermostable histone-like bacterial protein HUs significantly improves stability and DNA binding affinity.

Consensus-based protein engineering strategy has been applied to various proteins and it can lead to the design of proteins with enhanced biological performance. Histone-like HUs comprise a protein family with sequence variety within a highly conserved 3D-fold. HU function includes compacting and regulating bacterial DNA in a wide range of biological conditions in bacteria. To explore the possible impact of consensus-based design in the thermodynamic stability of HU proteins, the approach was applied using a dataset of sequences derived from a group of 40 mesostable, thermostable, and hyperthermostable HUs. The consensus-derived HU protein was named HUBest, since it is expected to perform best. The synthetic HU gene was overexpressed in E. coli and the recombinant protein was purified. Subsequently, HUBest was characterized concerning its correct folding and thermodynamic stability, as well as its ability to interact with plasmid DNA. A substantial increase in HUBest stability at high temperatures is observed. HUBest has significantly improved biological performance at ambience temperature, presenting very low Kd values for binding plasmid DNA as indicated from the Gibbs energy profile of HUBest. This Kd may be associated to conformational changes leading to decreased thermodynamic stability and, therefore, higher flexibility at ambient temperature.

Public health risk assessment associated with heavy metal and arsenic exposure near an abandoned mine (Kirki, Greece).

The 'Agios Philippos' lead-zinc mine in the Kirki region (NE Greece) is now closed, but its legacy of heavy metal contamination remains at the site. At present, management of the contaminated land is of major concern. The area is in a reclamation process and requires immediate remediation action, whereas human risks need to be carefully evaluated. In order to assess these risks, samples from around the mine were collected and analyzed and a scenario involving the oral, dermal, and inhaled doses of arsenic and heavy metals was formulated. A Monte Carlo approach was undertaken, in order to model the average daily dose and quantify the corresponding hazard index and cancer risk. A toxicological risk was associated with samples collected in the vicinity of the mine (floatation, mine tailings) and a pronounced carcinogenic risk for arsenic was evident at the broader occupational/environmental setting. These findings urge for immediate rehabilitation actions that will mitigate population exposures and promote long-term environmental safety in the area.