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一种基于超灵敏纳米纤维的纤维素酶促水解和深海微生物降解检测方法。

An ultrasensitive nanofiber-based assay for enzymatic hydrolysis and deep-sea microbial degradation of cellulose.

作者信息

Tsudome Mikiko, Tachioka Mikako, Miyazaki Masayuki, Uchimura Kohsuke, Tsuda Miwako, Takaki Yoshihiro, Deguchi Shigeru

机构信息

Research Center for Bioscience and Nanoscience, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka 237-0061, Japan.

SUGAR Program, JAMSTEC, 2-15 Natsushima-cho, Yokosuka 237-0061, Japan.

出版信息

iScience. 2022 Jul 30;25(8):104732. doi: 10.1016/j.isci.2022.104732. eCollection 2022 Aug 19.

Abstract

Substrates for enzymatic reactions, such as cellulose and chitin, are often insoluble in water. The enzymatic degradation of these abundant organic polymers plays a dominant role in the global carbon cycle and has tremendous technological importance in the production of bio-based chemicals. In addition, biodegradation of plastics is gaining wide attention. However, despite the significance, assaying these degradation reactions remains technically challenging owing to the low reaction rate, because only the surface of the substrate is accessible to the enzymes. We developed a nanofiber-based assay for the enzymatic hydrolysis of cellulose. This assay facilitated the quantification of the enzymatic hydrolysis of <1 ng crystalline cellulose. Utilization of the assay for the functional screening of cellulolytic microorganisms revealed an unprecedented genetic diversity underlying the production of deep-sea cellulase. This study reiterates that interdisciplinary efforts, such as from nanotechnology to microbiology, are critical for solving sustainability challenges.

摘要

酶促反应的底物,如纤维素和几丁质,通常不溶于水。这些丰富的有机聚合物的酶促降解在全球碳循环中起着主导作用,并且在生物基化学品的生产中具有巨大的技术重要性。此外,塑料的生物降解也受到广泛关注。然而,尽管其意义重大,但由于反应速率低,检测这些降解反应在技术上仍然具有挑战性,因为酶只能作用于底物的表面。我们开发了一种基于纳米纤维的纤维素酶促水解检测方法。该方法有助于对小于1纳克结晶纤维素的酶促水解进行定量。利用该检测方法对纤维素分解微生物进行功能筛选,揭示了深海纤维素酶产生背后前所未有的遗传多样性。这项研究重申,跨学科努力,如从纳米技术到微生物学,对于解决可持续发展挑战至关重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3367/9418596/67b2cf1c50bd/fx1.jpg

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