Higher order structure of Mucor miehei lipase and micelle size in cetyltrimethylammonium bromide reverse micellar system.
The higher order structure of Mucor miehei lipase and micelle size in a cationic cetyltrimethylammonium bromide (CTAB) reverse micellar system was investigated. Circular dichroic (CD) measurement revealed that the lipase far-UV CD spectra changed markedly, going from buffer solution to the reverse micellar solution, and were very similar for any organic solvent used. The ellipticity of the solubilized lipase in the far-UV region markedly decreased with increasing water content (W(0): molar ratio of water to CTAB), indicating that the secondary structure of lipase changed with the water content. The linear correlation between the W(0) and the micelle size was obtained by measuring dynamic light scattering. From the linear correlation between the micelle size and W(0), the higher order structure of the solubilized lipase appears to be affected directly by the micellar interface. The species and concentration of alcohol as a cosurfactant had an inferior effect on lipase structure. Especially, at ratios of 1-pentanol to CTAB of less than 8, the secondary and tertiary structures of lipase were preserved in the reverse micelles. The CTAB concentration had little effect on the lipase structure in the micelles. The catalytic activity of the lipase solubilized in the CTAB reverse micelles increased with increasing the W(0).
Naoe
Kazumitsu
K
Department of Chemical Engineering, Nara National College of Technology, 22 Yata, Yamato-Koriyama, Nara 639-1080, Japan. naoe@chem.nara-k.ac.jp
Takeuchi
Chihiro
C
Kawagoe
Mikio
M
Nagayama
Kazuhito
K
Imai
Masanao
M
D000438:Alcohols / Q000737:chemistry; D002384:Catalysis; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry*; D002942:Circular Dichroism; D008049:Lipase / Q000737:chemistry*; D008823:Micelles; D009089:Mucor / Q000201:enzymology*; D011487:Protein Conformation; D013056:Spectrophotometry, Ultraviolet; D013501:Surface-Active Agents / Q000737:chemistry
D000438:Alcohols; D002593:Cetrimonium Compounds; D008823:Micelles; D013501:Surface-Active Agents; D008049:Lipase; D000077286:Cetrimonium