Abstract:

 This study investigates the impact of various cosolvents on the activity and stability of two dihydrofolate reductases (DHFRs): bacterial R67 DHFR and human DHFR (hDHFR). The research addresses a significant gap in understanding how cosolvents influence enzyme behavior, particularly under conditions relevant to fragment-based drug design, which demands effective solubilization of test compounds without compromising enzyme function. The study’s primary aim is to identify cosolvents that maintain enzyme activity while facilitating the solubility of hydrophobic and hydrophilic compounds. The methodology involves assessing the effects of different cosolvents—methanol (MeOH), acetonitrile (ACN), dimethyl sulfoxide (DMSO), N,N-dimethylformamide (DMF), and imidazolium-based ionic liquids ([BMIm][BF4] and [BMIm][OctSO4])—on enzyme kinetics, thermal stability, and inhibition profiles.

The results show that MeOH and ACN retain the highest enzyme activity at moderate concentrations, while DMSO and DMF are more effective at solubilizing hydrophobic compounds but reduce enzyme activity more significantly. Ionic liquids show potential, but their high viscosity and lower solubilizing capacity limit their use. The main takeaway is that while MeOH offers the best compromise between enzyme activity retention and compound solubility, DMSO and DMF can be viable alternatives for early-stage screening of hydrophobic compounds, provided that their concentrations are carefully managed to balance solubility and enzyme activity​ (Bacteria – Escherichia …)​(Bacteria – Escherichia …)​(Bacteria – Escherichia …).

Conclusion on the Role of Constant Systems Cell Disruption equipment:

 Constant Systems Equipment plays a crucial role in this research by providing a consistent and controlled environment for conducting enzyme assays and thermal stability tests. This equipment ensures the reproducibility and reliability of the experimental conditions, which is essential for accurately assessing the impact of various cosolvents on the enzymes under study. By maintaining precise temperature and mixing conditions, the equipment helps to isolate the effects of the cosolvents from other variables, thereby enhancing the validity of the findings​ (Bacteria – Escherichia …)​(Bacteria – Escherichia …).

File Type: pdf
File Size: 310 KB
Categories: Bacteria, Enzymes, Escherichia coli
Constant Systems