Since
the introduction of deep eutectic solvents (DESs), numerous
reports have surfaced describing its tunable properties and environmentally
benign nature. Due to such favorable properties of DESs, they have
found a wide variety of applications. Moreover, in order to harness
the potential of proteins in numerous industries, there is an emergent
need to find a suitable cosolvent that is biocompatible with protein
and is also environmentally safe. In this context, this work presents
a systematic evaluation of effect of two deep eutectic solvents (DESs),
namely, choline chloride-urea (ChCl-urea) and choline chloride-glycerol
(ChCl-gly) on the structural and thermal stability along with activity
of enzyme α-chymotrypsin (CT) using circular dichroism (CD),
UV–visible, steady state, and thermal fluorescence spectroscopy.
It was observed that the presence of DESs does lead to enhancement
in the thermal stability of CT along with the preservation of activity.
The enzymatic activity was well maintained in both the DESs, and the
deleterious effect of urea was overcome by ChCl-urea on the enzyme.
Also, desirable results were observed for ChCl-urea, despite having
urea as one of its major components. Thus, the negative outcome of
urea was overpowered by the combination of ChCl and urea. Furthermore,
all the biomolecular studies were also performed with the individual
constituents of DESs. It was found that the effect imparted by both
the ChCl-based DESs on CT is by the virtue of DES itself rather than
its individual constituent. Overall, both the DESs can be described
as potential biocompatible, sustainable, and promising cosolvents
for CT with enhanced structural and thermal stability along with preservation
of its activity.
Proteins have immense untapped potential in numerous industries as a green catalyst. Thus, there is an emergent need to find a suitable co-solvent that is biocompatible with protein and environmentally...
Ionic liquids (ILs) represent as solvents or co-solvents for protein stabilization and refolding. Thus, ILs are replacement to toxic organic solvents in chemical, biotechnology and biomedical applications.
The present study demonstrated biological fluid inspired design of molecularly crowded IL media and disclosed an innovative and sustainable way for the packaging of Cyt c with enhanced activity and stability.
One of the major challenges in protein stability is that
proteins
can easily unfold in the presence of denaturants like urea, which
alters the native structure of proteins. There are numerous studies
in which ionic liquids (ILs) act as promising biocompatible solvents
(Bio-IL) for biomolecules. In this context, we present the refolding
ability of biocompatible imidazolium-based ILs, 1-ethyl 3-methyl imidazolium
ethyl sulfate [Emim][ESO4] (IL-1) and 1-ethyl 3-methyl
imidazolium chloride [Emim][Cl] (IL-2) against the chemically induced
structural changes in bovine and human serum albumin (BSA and HSA).
The work is substantiated with several spectroscopic, thermal and
docking studies. In steady-state fluorescence spectroscopy, we observe
that the emission intensity quenches for the protein in urea, which
is reversible with the addition of ILs. Circular dichroism spectral
studies reflect the reappearance of α helical content, which
is a good indicator of the refolding ability of ILs. Further, thermal
fluorescence studies showed that ILs have the ability to refold the
urea-induced denatured protein at a higher temperature range only
up to 7 M urea concentration; however, above 7 M urea concentration,
IL somehow fails to refold the protein. The work is also supported
by dynamic light scattering measurements, and the degree of BSA/HSA
aggregation was reduced with the introduction of Bio-IL to the urea–BSA/urea–HSA
system, ensuring the aggregate-free refolding. Furthermore, molecular
docking studies were employed to probe the binding sites, and the
results are well corroborated with the spectroscopic and thermal folding
results. Therefore, through this paper, we aim to unravel the mechanistic
intricacy of ILs using experimental and docking approaches. Overall,
ILs act as recoiling medium for both native and unfolded (denatured
by urea) BSA/HSA native structures.
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