Systematic determination of intrinsic reaction parameters in enzyme immobilizates
Systematic determination of intrinsic reaction parameters in enzyme immobilizates
复制标题
酶固定化物中固有反应参数的系统测定
DOI：
10.1016/j.ces.2009.12.026
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发表时间：
2010
影响因子：
4.7
通讯作者：
Spiess AC
中科院分区：
文献类型：
--
作者：
Zavrel M;Michalik C;Schwendt T;Schmidt T;Ansorge-Schumacher M;Janzen C;Marquardt W;Büchs J;Spiess AC
为了合理地设计固定化酶的过程，需要一个机械动力学模型，该模型考虑了所有的动力学和热力学现象，包括酶反应、反应物在两相之间的传质以及它们在固定化物中的扩散。以有机溶剂中悬浮的水凝胶小球为例，采用基于模型的实验分析方法，得到了酶固定化的机理动力学模型。结果表明，通常采用的体相浓度测量方法不足以得出机械性结论。发现最合适的测量技术是沿着水凝胶珠半径的浓度的量化。这些由双光子激光扫描显微镜实现的线扫描首次允许同时估计本征反应和传质参数。因此，所获得的两相水凝胶体系的本征参数估计值可以直接与在各个体系中所获得的估计值进行比较。这一比较首次表明，温和的水凝胶包裹对酶反应没有显著影响。然而，发现了对传输参数的重大影响，这突显了使用机械模型分析真实反应系统的必要性。
For the rational design of processes using immobilized enzymes a mechanistic kinetic model is required, which accounts for all kinetic and thermodynamic phenomena, including the enzyme reaction, the mass transfer of the reactants between both phases, and their diffusion inside the immobilizate. For the example of enzymes immobilized in hydrogel beads suspended in an organic solvent, such a mechanistic kinetic model was obtained by a model-based experimental analysis approach. It was proven that the usually applied concentration measurements in the bulk phase are not sufficient to draw mechanistic conclusions. The most suitable measurement technique was found to be the quantification of the concentration along the radius of the hydrogel bead. These line scans, achieved by two-photon laser scanning microscopy, for the first time allowed to estimate intrinsic reaction and mass transfer parameters simultaneously. Thus, the obtained intrinsic parameter estimates for the biphasic hydrogel system could be directly compared with those obtained in individual systems. This comparison revealed for the first time that the enzyme reaction was not significantly affected by the mild hydrogel encapsulation. However, a significant impact on the transport parameters was found that underlines the need for analyzing the real reaction system using mechanistic models.
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