Surface Pitting on Nanofibrous Matrix as an Ultra- Sensitive Indicator for Enzymatic Hydrolysis of Crystalline Cellulose
Surface Pitting on Nanofibrous Matrix as an Ultra- Sensitive Indicator for Enzymatic Hydrolysis of Crystalline Cellulose
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DOI：
10.26434/chemrxiv.14562654
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发表时间：
2021-05
期刊：
影响因子：
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通讯作者：
Mikiko Tsudome;M. Tachioka;Miwako Tsuda;Y. Takaki;S. Deguchi
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文献类型：
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作者：
Mikiko Tsudome;M. Tachioka;Miwako Tsuda;Y. Takaki;S. Deguchi
测定不溶于水的底物（例如纤维素和合成聚合物）的酶促降解在技术上仍然具有挑战性，主要是因为酶只能接触到底物的表面，并且与水溶性底物相比，反应进行得非常缓慢。在这里，我们展示了纤维素酶水解的超灵敏半定量测定。通过将纳米纤维基质与压电驱动喷墨打印和光学轮廓测定法相结合，成功量化了小于 1 纳克结晶纤维素的酶水解。当应用相同的原理来阐明深海中微生物对纤维素的降解时，揭示了纤维素酶前所未有的遗传多样性。这项工作表明，真正的跨学科努力，涵盖从纳米技术到微生物学的不同学科，对于解决可持续发展的科学和技术问题至关重要。
Assaying enzymatic degradation of the water-insoluble substrate such as cellulose and synthetic polymers has remained technically challenging, primarily because only the surface of the substrate is accessible to the enzymes and the reaction proceeds very slowly compared with those of water-soluble substrates. Here we show an ultra-sensitive and semi-quantitative assay for enzymatic hydrolysis of cellulose. By combining nanofibrous matrices with piezo-driven inkjet printing and optical profilometry, enzymatic hydrolysis of less than 1 nanogram of crystalline cellulose was successfully quantified. Unprecedented genetic diversity of cellulase was revealed when the same principle was applied for elucidating microbial degradation of cellulose in the deep sea. This work demonstrates that truly interdisciplinary efforts, encompassing diverse disciplines from nanotechnology to microbiology, are crucial to address scientific and technological problems towards sustainability.