The integration of enzymes with solid materials is important in many biotechnological applications, including the use of immobilized enzymes for biocatalytic synthesis. The development of functional enzyme-material composites is restrained by the lack of molecular-level insight into the behavior of enzymes in confined, surface-near environments. Here, we review recent advances in surface-sensitive spectroscopic techniques that push boundaries for the determination of enzyme structure and orientation at the solid-liquid interface. We discuss recent evidence from single-molecule studies showing that analyses sensitive to the temporal and spatial heterogeneities in immobilized enzymes can succeed in disentangling the effects of conformational stability and active-site accessibility on activity. Different immobilization methods involve distinct trade-off between these effects, thus emphasizing the need for a holistic (systems) view of immobilized enzymes for the rational development of practical biocatalysts.

中文翻译：

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关于固体表面固定化酶的结构与催化效率之间的关系：方法学的进步和对单分子观点的追求。

酶与固体材料的整合在许多生物技术应用中都很重要，包括使用固定化酶进行生物催化合成。功能性酶材料复合材料的发展受到缺乏分子水平的洞察力的限制，这些洞察力仅限于在狭窄的，近表面环境中使用的酶的行为。在这里，我们回顾了表面灵敏光谱技术的最新进展，这些技术为确定酶在固液界面的结构和方向提供了便利。我们讨论了单分子研究的最新证据，这些研究表明，对固定化酶的时空异质性敏感的分析可以成功地解开构象稳定性和活性位点可及性对活性的影响。