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Simple Protocol to Facilitate Students’ Understanding of the Effects of Enzyme Immobilization on Kinetics of Reaction and Mass Transfer
Journal of Chemical Education ( IF 2.5 ) Pub Date : 2020-07-21 , DOI: 10.1021/acs.jchemed.0c00848 Manuel Simões 1 , F. Xavier Malcata 1
Journal of Chemical Education ( IF 2.5 ) Pub Date : 2020-07-21 , DOI: 10.1021/acs.jchemed.0c00848 Manuel Simões 1 , F. Xavier Malcata 1
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A laboratory experiment is described in which students carry out immobilization of the enzyme invertase via entrapment in alginate (Ca-alginate) and, accordingly, estimate values for kinetic constants and mass transfer coefficients. Immobilization of enzymes, or confinement thereof to a defined space with retention of most catalytic activity, is attractive for allowing repeated (and continuous) use. However, interest in immobilization hinges critically on whether and how it affects kinetic performance. The overall goal is indeed to retain as much activity as possible and minimize alteration of selectivity, while extending its useful life for as long as feasible. Toward this goal, it is relevant to quantify the effects of immobilization upon enzyme kinetics and mass transfer in particular—with the latter constraining observed (rather than intrinsic) reaction kinetics, owing to diffusional resistance. Invertase was selected as a model enzyme since the associated laboratory handling is safe, fast, reproducible, not expensive, and prone to easily attain the intended learning outcomes.
中文翻译：
简单的协议，以促进学生了解酶固定化对反应动力学和传质动力学的影响
描述了一个实验室实验，在该实验中，学生通过诱捕藻酸盐（Ca-藻酸盐）来固定酶转化酶，并据此估算动力学常数和传质系数的值。酶的固定化或将其限制在限定的空间内并保留最大的催化活性，对于允许重复（和连续）使用是有吸引力的。然而，对固定化的兴趣关键取决于它是否以及如何影响动力学性能。实际上，总体目标是保留尽可能多的活性并最大程度地减少选择性的变化，同时尽可能延长其使用寿命。为了这个目标，特别重要的是要量化固定化对酶动力学和质量转移的影响，尤其是由于扩散阻力，后者会限制（而不是固有的）反应动力学。选择蔗糖酶作为模型酶，因为相关的实验室操作安全，快速，可重现，不昂贵且易于获得预期的学习成果。
更新日期：2020-08-11
中文翻译：
简单的协议，以促进学生了解酶固定化对反应动力学和传质动力学的影响
描述了一个实验室实验，在该实验中，学生通过诱捕藻酸盐（Ca-藻酸盐）来固定酶转化酶，并据此估算动力学常数和传质系数的值。酶的固定化或将其限制在限定的空间内并保留最大的催化活性，对于允许重复（和连续）使用是有吸引力的。然而，对固定化的兴趣关键取决于它是否以及如何影响动力学性能。实际上，总体目标是保留尽可能多的活性并最大程度地减少选择性的变化，同时尽可能延长其使用寿命。为了这个目标，特别重要的是要量化固定化对酶动力学和质量转移的影响，尤其是由于扩散阻力，后者会限制（而不是固有的）反应动力学。选择蔗糖酶作为模型酶，因为相关的实验室操作安全，快速，可重现，不昂贵且易于获得预期的学习成果。
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