Enhanced Catalytic Activity of α-Chymotrypsin in Cationic Surfactant Solutions: The Component Specificity Revisited.
Enhanced catalytic activity (super activity) of enzymes in the presence of surfactants is of key importance in "micellar enzymology"; such super activity is not very trivial, it is highly system specific, and the mechanism behind the activity enhancement is not always well apprehended. We report the catalytic activity of α-chymotrypsin (CHT) on ala-ala-phe-7-amido-4-methylcoumarin (AMC) in the presence of cationic surfactants of different hydrophobic chain lengths: dodecyltrimethylammonium bromide (DTAB), cetyltrimethylammonium bromide (CTAB) and octadecyltrimethylammonium bromide (OTAB). It is observed that in comparison to buffer the catalytic activity of CHT is enhanced 5-fold in premicellar DTAB solutions, while negligible changes are observed in CTAB and OTAB. Activity decreases considerably in the post micellar concentration, specifically for the latter two surfactants. A similar trend is also obtained in another substrate 2-napthyal acetate hydrolysis. Such surfactant specific superactivity is intriguing. The protein's secondary and tertiary structures in the presence of these surfactants are determined using circular dichroism (CD) spectroscopy and it is found that both CTAB and OTAB perturb the protein structure significantly, especially in the post micellar concentrations. DTAB, on the other hand, does not produce noticeable changes in the protein structure. The various pairwise interactions present in the system have been underlined using both steady-state and time-resolved fluorescence spectroscopy. Assuming a three-step kinetics model, we determine the free energy changes of the reaction, and the observations have been discussed in the light of the various interactions among the components.
Department of Chemical, Biological and Macromolecular Sciences, S. N. Bose National Centre for Basic Sciences , Block JD, Sector III, Salt Lake, Kolkata 700106, India.
Animesh
A
Patra
Department of Chemical, Biological and Macromolecular Sciences, S. N. Bose National Centre for Basic Sciences , Block JD, Sector III, Salt Lake, Kolkata 700106, India.
Nirnay
N
Samanta
Department of Chemical, Biological and Macromolecular Sciences, S. N. Bose National Centre for Basic Sciences , Block JD, Sector III, Salt Lake, Kolkata 700106, India.
Dipak Kumar
DK
Das
Department of Chemical, Biological and Macromolecular Sciences, S. N. Bose National Centre for Basic Sciences , Block JD, Sector III, Salt Lake, Kolkata 700106, India.
Rajib Kumar
0000-0001-9159-0517
RK
Mitra
D055162:Biocatalysis / Q000187:drug effects; D002918:Chymotrypsin / Q000737:chemistry*; D003374:Coumarins / Q000737:chemistry; D004735:Energy Transfer; D008823:Micelles; D015394:Molecular Structure; D009842:Oligopeptides / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D013501:Surface-Active Agents / Q000737:chemistry*
D003374:Coumarins; D008823:Micelles; D009842:Oligopeptides; D000644:Quaternary Ammonium Compounds; D013501:Surface-Active Agents; C117513:alanyl-alanyl-phenylalanyl-7-amino-4-methylcoumarin; D002918:Chymotrypsin; C117608:alpha-chymotrypsin