Naoe Kazumitsu, Takeuchi Chihiro, Kawagoe Mikio, Nagayama Kazuhito, Imai Masanao
Department of Chemical Engineering, Nara National College of Technology, 22 Yata, Yamato-Koriyama, Nara 639-1080, Japan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 May 1;850(1-2):277-84. doi: 10.1016/j.jchromb.2006.11.048. Epub 2006 Dec 13.
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).
研究了米黑根毛霉脂肪酶在阳离子十六烷基三甲基溴化铵(CTAB)反胶束体系中的高阶结构和胶束尺寸。圆二色性(CD)测量表明,从缓冲溶液到反胶束溶液,脂肪酶的远紫外CD光谱发生了显著变化,并且对于所使用的任何有机溶剂来说都非常相似。随着水含量(W(0):水与CTAB的摩尔比)增加,溶解在反胶束中的脂肪酶在远紫外区域的椭圆率显著降低,这表明脂肪酶的二级结构随水含量而变化。通过测量动态光散射获得了W(0)与胶束尺寸之间的线性相关性。从胶束尺寸与W(0)之间的线性相关性来看,溶解的脂肪酶的高阶结构似乎直接受到胶束界面的影响。作为助表面活性剂的醇的种类和浓度对脂肪酶结构的影响较小。特别是,当1-戊醇与CTAB的比例小于8时,脂肪酶的二级和三级结构在反胶束中得以保留。CTAB浓度对胶束中脂肪酶的结构影响很小。溶解在CTAB反胶束中的脂肪酶的催化活性随着W(0)的增加而提高。