Comparison of the different types of surfactants for the effect on activity and structure of soybean peroxidase.
In the pH 2.6 and 5.2 systems, soybean peroxidase (SBP) (isoelectric point, pI 3.9) has positive and negative charge, respectively. In order to acquire detailed knowledge on the role played by electrostatics in the denaturation of proteins, a comparison of anionic surfactant sodium dodecyl sulfate (SDS), nonionic surfactant nonaethylene glycol monododecyl ether [C12H25O(CH2CH2O)9H] (AEO9), and cationic surfactant cetyltrimethylammonium bromide (CTAB) for the influences on the activity and structure of soybean peroxidase (SBP) was carried out by measuring the activity, far-UV circular dichrosm, fluorescence, and electronic absorption spectra of SBP in the pH 2.6 and 5.2 systems at 30 degrees C. In the pH 2.6 systems, the interaction of SDS with SBP results in an increase in the fluorescence intensity with a red shift of the emission maximum of the tryptophan fluorescence and a blue shift of the Soret band. In the meantime, the alpha-helix of SBP is unfolded and the activity of SBP is lost irreversibly. In pH 5.2 systems, the fluorescence spectra features of SBP are similar to those in pH 2.6 systems with increasing SDS concentration, but a red shift of Soret band as well as an alteration of the tertiary structure of SBP occurs, and the lost activity is recoverable. The electrostatic interactions between SBP and SDS play an important role in the denaturation of SBP. The effects of AEO9 and CTAB in pH 2.6 and 5.2 systems on the activity and spectral features of SBP are similar to that of SDS in pH 5.2 systems, but AEO9 is prone to unfold the beta-sheet of SBP in pH 2.6 systems. The electrostatic interactions of CTAB with SBP are not the primary elements for denaturation of SBP, which distinctly differ from those of SDS. These results can be useful with respect to wide applications of the surfactants in the separation and purification of proteins.
Zhang
Weican
W
State Key Laboratory of Microbial Technology, Shandong University, Jinan, Shandong, China, 250100.
Dai
Xuhui
X
Zhao
Yue
Y
Lu
Xuemei
X
Gao
Peiji
P
D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D009195:Peroxidase / Q000378:metabolism*; D000077423:Polidocanol; D011092:Polyethylene Glycols / Q000737:chemistry / Q000494:pharmacology; D017433:Protein Structure, Secondary; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry / Q000494:pharmacology; D013025:Glycine max / Q000187:drug effects* / Q000201:enzymology*; D013501:Surface-Active Agents / Q000737:chemistry* / Q000494:pharmacology*
D002593:Cetrimonium Compounds; D013501:Surface-Active Agents; D000077423:Polidocanol; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D009195:Peroxidase; D000077286:Cetrimonium