Mechanism of deep eutectic solvents enhancing catalytic function of cytochrome P450 enzymes in biosynthesis and organic synthesis

Current understanding and insights towards protein stabilization and activation in deep eutectic solvents as sustainable solvent media

“…The hydrogen-bonding network present in the DES can be utilized as a protective layer to eliminate any ion from penetrating the helical structure, thus providing stabilization. 123 In accordance with the above mentioned mechanism, the simulation results displayed that the TMG–ethylene glycol DES completely unfolds the protein, while the protein slightly unfolds in the case of the TMG–glycerol DES. This is ascribed to the stronger hydrogen bonding between TMG–glycerol than TMG–ethylene glycol; hence, more penetration into the protein core occurs in the case of TMG–ethylene glycol, which leads to protein structural disruption.…”

“…Furthermore, Yao's research group 123 revealed the structural changes of cytochrome-P450 BM-3 induced in the presence of ChCl–urea DES containing different water contents. It was observed that the enzyme was reversible towards thermal unfolding in the presence of DES.…”

“…This study also uncovered that the enzyme displayed high long term thermal stability in the alcohol-based DES. 123…”

“…This was confirmed through the values of docking affinity energies, which depend on the number of hydrogen bonds with enzyme AAs. Furthermore, Sun et al 123 also mentioned that the hydrogen bond networks induced by DES could be utilized as a protective layer against ions to interfere in the helical structure of the protein domain, consequently stabilizing protein. Inspired by the above findings, recently our group has demonstrated the mechanism of protein stabilization via the formation of a hydration shell around the active site of the protein.…”

Deep eutectic solvents as a stabilising medium for NAD coenzyme: unravelling the mechanism behind coenzyme stabilisation effect

“…Additionally, all DES–buffer systems tested (50 : 50) had a very pronounced stabilising effect on peroxygenase (much greater compared to referent buffer). Sun et al 72 investigated the catalytic activity and thermal stability of NAD-dependent cytochrome P450 in a series of ChCl-based DES paired with urea and polyols (such as Gly, EG and PG) and observed that both the activity and the stability of the enzymes in DES were higher than that in buffer solution. For example, in ChCl:U at water contents above 60% improved enzyme catalytic activity and stability compared to reference organic solvent was observed (activity at lower water contents was not monitored).…”