Tunable Semicrystalline Thin Film Cellulose Substrate for High-Resolution, In-Situ AFM Characterization of Enzymatic Cellulose Degradation
Tunable Semicrystalline Thin Film Cellulose Substrate for High-Resolution, In-Situ AFM Characterization of Enzymatic Cellulose Degradation
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DOI：
10.1021/acsami.5b09948
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发表时间：
2015-12-23
影响因子：
9.5
通讯作者：
Plank, Harald
中科院分区：
文献类型：
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作者：
Ganner, Thomas;Rosker, Stephanie;Plank, Harald
在酶促纤维素降解领域，不同酶与多态纤维素材料之间的基本相互作用至关重要，但尚未完全了解。原子力显微镜（AFM）是一种具有直接可视化这些生物过程潜力的技术，因为它具有实时的原位能力；从空间分辨率到分子尺度的研究。为了充分利用该技术的能力并揭示基本的酶-纤维素生物过程，合适的纤维素底物是决定性的。在这项研究中，我们介绍了一种半晶薄膜纤维素（SCFTC）底物，它通过以下方式满足了对这种理想纤维素底物的强烈要求：(1)可调节的多态性：通过在无定形纤维素基质中嵌入不同含量的均匀大小的纤维素纳米晶体；(2)用于高分辨率、高速AFM的纳米平面拓扑；(3)快速、简单、可重复制造。该研究从SCTFC制备方案的详细描述开始，包括深入的材料表征。在第二部分中，我们通过联合、单独和顺序暴露于TrCel6A/TrCel7A纤维素酶（里氏木霉）来证明SCTFC底物在酶降解研究中的适用性，以可视化纳米级的协同效应。
In the field of enzymatic cellulose degradation, fundamental interactions between different enzymes and polymorphic cellulose materials are of essential importance but still not understood in full detail. One technology with the potential of direct visualization of such bioprocesses is atomic force microscopy (AFM) due to its capability of real-time in situ;investigations with spatial resolutions down to the molecular scale. To,exploit the full capabilities of this technology and unravel fundamental enzyme-cellulose bioprocesses, appropriate Cellulose substrates are decisive. In this study, we introduce a semicrystalline-thin-film-cellulose (SCFTC) substrate which fulfills the strong demands on such ideal cellulose substrates by means of (1) tunable polymorphism via:Variable contents of homogeneously sized cellulose nanocrystals embedded in an amorphous cellulose matrix; (2) nanoflat surface topology for high-resolution and high-speed AFM; and (3) fast, simple, and reproducible fabrication. The study starts with a detailed description of SCTFC preparation protocols including an in-depth material characterization. In the second part, we demonstrate the suitability of SCTFC substrates for enzymatic degradation studies by combined, individual, and sequential exposure to TrCel6A/TrCel7A cellulases (Trichoderma reesei) to visualize synergistic effects down to the nanoscale.