Pastorino Laura, Disawal Sandeep, Nicolini Claudio, Lvov Yuri M, Erokhin Victor V
Department of Biophysical M&O Science and Technologies, University of Genoa, Corso Europa 30, 16132 Genoa, Italy.
Biotechnol Bioeng. 2003 Nov 5;84(3):286-91. doi: 10.1002/bit.10761.
In the present work the layer-by-layer nano-assembly technique was used for the development of complex catalytic microparticles on the basis of firefly luciferase (FL). FL films containing 1, 2, or 3 monolayers were assembled on silver electrode QCM-resonators and on 520-nm diameter sulfonated polystyrene latex by alternate adsorption of FL and polycations using electrostatic interactions for the interlayer interaction. The assembly process was studied with quartz crystal microbalance, UV-vis spectroscopy, and microelectrophoresis (surface potential). Structural studies of the resulting multilayers confirmed stepwise deposition of FL and cationic poly(dimethyldiallyl ammonium chloride) with a bilayer thickness of 14 nm; a systematic shift of the surface potential from +28 mV for poly(dimethyldiallyl ammonium chloride) to -14 mV for luciferase outermost layer was established. The functionality and stability of the biocolloids were demonstrated by monitoring the intensity of the light emission. Factors influencing the light emitted upon catalytic activity of FL such as the number of luciferase layers in the film and polyion layer at the outermost layer were studied.
在本工作中，基于萤火虫荧光素酶（FL），采用层层纳米组装技术来制备复合催化微粒。通过利用静电相互作用进行层间相互作用，使 FL 和聚阳离子交替吸附，在银电极 QCM 谐振器以及直径为 520nm 的磺化聚苯乙烯胶乳上组装了含有 1、2 或 3 个单层的 FL 薄膜。利用石英晶体微天平、紫外可见光谱和微电泳（表面电位）研究了组装过程。对所得多层结构的研究证实了 FL 和阳离子聚（二甲基二烯丙基氯化铵）的逐步沉积，双层厚度为 14nm；确定了表面电位从聚（二甲基二烯丙基氯化铵）的 +28mV 到荧光素酶最外层的 -14mV 的系统性变化。通过监测发光强度证明了生物胶体的功能和稳定性。研究了影响 FL 催化活性时发光的因素，如薄膜中荧光素酶层数和最外层聚离子层等。
