Imidazolium bromide-based ionic liquid assisted improved activity of trypsin in cationic reverse micelles.
The present work reports the imidazolium-based ionic liquids (ILs) assisted enhancement in activity of water-pool solubilized enzyme trypsin in cationic reverse micelles of CTAB. A set of imidazolium ILs (1-alkyl-3-methyl imidazolium bromides) were prepared with varying lengths of their side arm which results in the differential location of these organic salts in the reverse micelles. The different ILs offered varied activating effects on the biocatalyst. The activity of trypsin improved approximately 30-300% in the presence of 0.1-10 mM of different ILs in reverse micelles of CTAB. Trypsin showed approximately 300% (4-fold) increment in its activity in the presence of IL 2 (1-ethyl-3-methyl imidazolium bromide, EMIMBr) compared to that observed in the absence of IL in CTAB reverse micelles. The imidazolium moiety of the IL, resembling the histidine amino acid component of the catalytic triad of hydrolases and its Br(-) counterion, presumably increases the nucleophilicity of water in the vicinity of the enzyme by forming a hydrogen bond that facilitates the enzyme-catalyzed hydrolysis of the ester. However, the ILs with increasing amphiphilic character had little to no effect on the activity of trypsin due to their increased distance from the biocatalyst, as they tend to get localized toward the interfacial region of the aggregates. Dynamic light scattering experimentation was carried out in the presence of ILs to find a possible correlation between the trypsin activity and the size of the aggregates. In concurrence with the observed highest activity in the presence of IL 2, the circular dichroism (CD) spectrum of trypsin in CTAB reverse micelles doped with IL 2 exhibited the lowest mean residue ellipticity (MRE), which is closest to that of the native protein in aqueous buffer.
Department of Biological Chemistry, Indian Association for the Cultivation of Science Jadavpur, Kolkata-700 032, India.
Sisir
S
Debnath
Dibyendu
D
Das
Sounak
S
Dutta
Prasanta Kumar
PK
Das
D002412:Cations; D000077286:Cetrimonium; D002593:Cetrimonium Compounds; D002942:Circular Dichroism; D006868:Hydrolysis; D007093:Imidazoles / Q000737:chemistry*; D007477:Ions; D009682:Magnetic Resonance Spectroscopy; D008823:Micelles; D013056:Spectrophotometry, Ultraviolet; D014357:Trypsin / Q000737:chemistry*
D002412:Cations; D002593:Cetrimonium Compounds; D007093:Imidazoles; D007477:Ions; D008823:Micelles; D014357:Trypsin; D000077286:Cetrimonium